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The genus Melocactus (“cabeça-de-frade”) comprises 32 species in Brazil, of which M. glaucescens and M. paucispinus are threatened with extinction. The present work evaluated the effects of different concentrations of Murashige & Skoog (MS, MS/2 and MS/4) culture medium and sucrose (15 g L-1 and 30 g L-1) on in vitro seed germination and plant growth of M. glaucescens and the efficiency of sterilization with sodium hypochlorite (NaOCl) on in vitro seed germination and plant growth of M. glaucescens when using seeds and M. glaucescens and M. paucispinus when using apical segment of cladode. In M. glaucescens, the final germination at the different MS and sucrose concentrations varied between 53.5 and 68.1% and the best results for in vitro growth were observed with the lowest mineral salt (MS/2 and MS/4) and sucrose (15 g L-1) concentrations, with lengths of the aerial portion of 9.70 and 10.76 mm, respectively. There was no difference in seed germination and plant growth in chemical and autoclave medium. It is concluded that the use of chemical sterilization with NaOCl at low concentrations of salts (MS/2 and MS/4) and sucrose (15 g L-1) are quite advantageous for producing ornamental plants germinated in vitro and/or apical segment of cladode of M. glaucescens andM. paucispinus, representing a reduction of costs for in vitro cultivation of this species.

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The high endemism, the natural habitat degradation, and the over-collection for ornamental purposes have led some species such as Melocactus paucispinus and M. glaucescens to be threatened with extinction. The use of in vitro conservation techniques, such as slow growth storage, promotes the preservation of genetic diversity with integrity. The goal of this study was to establish a strategy for in vitro conservation of apical segments of the cladode of M. paucispinus and M. glaucescens and evaluate the genetic diversity of individuals from in vitro germinated plants. For such purpose, different concentrations of the plant regulator ancymidol and the osmotic agent sucrose on the inhibition of the in vitro growth were tested, and the genetic diversity of M. paucispinus and M. glaucescens individuals stored in vitro was evaluated. Sucrose showed higher efficiency in the reduction of growth than ancymidol for both species. However, due to the reduction in survival percentage, the use of sucrose over 75 g L− 1 in the in vitro conservation of both species for 360 days is not recommended. In the genetic diversity analysis, 76.92% of polymorphic loci (P), expected heterozygosity (He) = 0.276 and Shannon index (S) = 0.414 were observed for M. paucispinus. For M. glaucescens, the observed values were P = 95.38%, He = 0.228 and S = 0.369. These values observed here were higher than those previously found for the natural populations of these species, which demonstrated that this in vitro collection showed genetic diversity and can be used in management and reintroduction programs of these species.